• Korean Journal of Ichthyology
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• v.34 no.2
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• pp.86-95
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• 2022

Egg development and morphological change of larvae and juvenile of the Roughscale sole, Clidoderma asperrimum, were investigated in the present study. Adult fishes were collected on the East Sea, Korea, from 2017 to 2018 and reared in a circular water tank (Ø 6×1 m) at water temperature of 12.8±1.9℃. Fertilized eggs ranged from 1.42 to 1.59 mm (mean 1.51±0.04 mm, n=50) in diameter. The eggs were spherical in shape, transparent, floating and colorless. The egg yolk was separated from the egg membrane 60 mins post-fertilization (PF), and an embryo was formed in 62 hrs PF. More than 50% of the eggs hatched within 144 hrs PF in the range of 10.2~11℃(mean 10.8℃). The size of the newly hatched larvae were 4.22~4.64 mm (mean 4.53±0.16 mm) in total length (TL), their mouth and anus were not open yet. At 10 days after hatching, the preflexion larvae reached 5.88~6.62 mm (mean 6.31±0.33 mm) in TL, and the yolk absorption was completed and the mouth began to open. At 55 days after hatching the larvae reached to flexion larvae stage and they were 10.4~13.3 mm (mean 12.7±1.3 mm) in TL, and the tip of notochord was bent upward. At 120 days after hatching the larvae reached to juvenile stage and they were 35.3~40.5 mm (mean 39.5±2.4 mm) in TL. Their all fins had completed set of the fin-rays (D. 79~94: A. 63~75) and the juveniles adopted a benthic life.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.30-30
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• 2001

The culture of preantral follicles has important biotechnological implications through its potential to produce the large quantity of oocytes for embryo production, transgenesis research, conservation of rare breed, and a potential source of ovarian genetic material. The present study was conducted to establish the optimal conditions of in vitro culture for intact bovine preantral follicles; and to examine the developmental ability of oocytes derived from the in vitro-grown preantral follicles; and to investigate the effects of various concentrations of FSH and LH on these processes. Bovine preantral follicles (150 $\pm$ 1.2${\mu}{\textrm}{m}$), surrounded by theca cell, were isolated enzymetically and mechanically from ovarian cortical slides in Leibovitz L-15 medium containing 1 mg/$m\ell$ collagens and 0.2 mg/$m\ell$ DNase I and cultured for 25 days in the presence of different concentrations of bovine FSH and LH in $\alpha$MEM medium with insulin, transferrin, and selenite. The survival was tested by frypan Blue and Hematoxylin. The survival and growth rates of follicles were higher in FSH treatment groups than these in control (P<0.001), but there were no significant differences between the LH treatment groups and the control. In 25 days, the survival and growth rates of follicles in FSH and LH treatment group (50%, 300$\pm$1.0${\mu}{\textrm}{m}$) were higher than in FSH treatment group (40%, 244$\pm$0.5${\mu}{\textrm}{m}$) and the control group (25%, 160$\pm$ 1.0${\mu}{\textrm}{m}$). Fifty-five percent of healthy antral follicles were obtained, and 60% of the oocytes complete meiotic maturation to the metaphase II stage. Twenty-two percent of the mature oocytes underwent cleavage, and 9% developed to the blastocyst stage. In this study, in vitro-grown oocytes (111 $\pm$ $1.5mutextrm{m}$), under our culture conditions, were not equivalent in size to the in vivo-grown oocytes (130$\pm$1.3${\mu}{\textrm}{m}$). Therefore, these results suggest that bovine preantral follicles with intact theca cell can grow to the antral stage in 25days, and that oocytes from those follicles can acquire the meiotic competence and normally undergo fertilization and development to the blastocyst stage. However, the developmental capacity of in vitro-grown oocytes is presumably not comparable to those of the in vivo counterparts.

• Journal of Practical Agriculture & Fisheries Research
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• v.20 no.2
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• pp.89-97
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• 2018

This study was conducted to elucidate the effect of after-ripening of gourd fruits before seed collection. Five varieties of commercial gourd varieties were cultivated and the fruits were picked after 24, 31, 50, and 70 days from pollination. For after-ripening treatments the fruits were placed for five days in a cool, air-circulated place while a number of fruits were placed for longer periods to 85 days including growing period, 24 + 62 days, 31 + 55 days, 50 + 35 days, and 70 + 15 days. The seeds collected from fruits harvested on 24^th day were not emerged at all, but the seeds from after-ripening treatment for 62 days showed 78% emergence or higher. The growth of seedlings showed the effects of after-ripening treatment of the fruits before seed collection; the seeds collected after ripening treatment showed better hypocotyl growth and larger embryo size than the non-treated seeds.

• Korean Journal of Plant Resources
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• v.15 no.3
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• pp.177-187
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• 2002

In our experiment, selected mutants were used which showed not only the phenotype of a specific unpolished rice but also phenotypes of EM 40, LO 1050, and TAL 214. Reciprocal crosses between the mutants were conducted to select strains which would have more quantity of lipids than before. The constitution of fatty acid was also tested to figure out nutritional aspects of the mutants. In the crossing between EM 40 mutants and mutants (LO 1050) having a thick aleurone layer, the expression of EM 40 mutants has no relation with the thickness of the aleurone layer. And the lipid content of new F$_2$ strains through the crossing is 4.15 ％. The lipid content is larger than those of the parents including Kinmaze and in other crossings of this experiment. This is attributed to the fact that the new F$_2$ strains are the products of the crossing between genes responsible for the size of buds, where lipid is accumulated, and genes accountable for the thickness of the aleurone layer. In the crossing between EM 40 mutants and TAL 214 mutants, lipid content of the new F$_2$ strains is 3.8 ％, higher than 2.92 ％ of TAL 214 mutants. But the degree of lipid increase is smaller than in two other crossings. This is probably because genes expressing the phenotypes of TAL 214 affect the size of EM 40, which gets smaller. The aleurone layer of the new F$_2$ strains is 12 $\mu\textrm{m}$ thicker than the layer of TAL 214 mutants, but 6 $\mu\textrm{m}$ thinner than that of parents (LO 1050) having a thick aleurone layer. This seems to be affected by the size of a microscope. The phenotype of the new F$_2$ strains appears to be similar to that of TAL 214. The lipid content of the new F$_2$ strains is 3.85 ％, larger than 2.92 ％ of TAL 214 and 3.01 % of LO 1050. The increase may be due to the aleurone layer of LO 1050. And the size of the bud of the unpolished rice, though it is not big enough like that of LO 1050, seems to be affected by the accumulation of genes in the thick aleurone layer. The accumulation may contribute to the increase in the content of lipid. When it comes to the constitution of fatty acid, there is little difference between parents like Kinmaze and the new F$_2$ strains. But oleic acid increases while linoleic acid decreases. And the decrease in the linolenic acid seems to contribute to the increase in lipid content. This fact also raises the possibility that genes accountable for specific phenotypes could change the quality of rice if the genes are accumulated. Now, experiments on strains which have large lipid content in EM 40 type 1(ge-1, 3.68 ％), EM type 2(ge-2, 2.91 ％), thick aleurone layer(4.63 ％), and starch layer(3.44 ％) are under way to figure out the effects of gene accumulation. These experiments are likely to present the ways for increasing the lipid content.

• The Korean Journal of Malacology
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• v.23 no.2
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• pp.189-198
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• 2007

The reproductive cycle, egg capsules in the egg-mass, first sexual maturity, and sex ratio of the bladder moon, Glossaulax didyma ($R\ddot{o}ding$) were investigated. The gastropods collected from the intertidal zone of Biin Bay, Seocheon, Korea were studied by using histological analysis and morphometric data. The gonadosomatic index (GSI) of females and males began to increase in March and reached maximum in May. Then their values sharply decreased from late in May to August due to spawning. The condition index (CI) began to increase in February and reached maximum in May, then gradually declined in the spawning period. The CI calculated for determination of the spawning period was coincided with changes in the GSI and gonadal phases. Spawning occurred between late in May to August in females and early in May to August in males. Spawning peak was observed between July and August when the seawater temperature rose to 19 $^{\circ}C$. Reproductive cycle with the gonadal development phases of this species can be divided into five successive stages in females and four in males: in females, early active stage (December to February), late active stage (February to March), ripe stage (April recovery stage (August to November); in males, active stage (December to March), ripe stage (March to July), copulation stage (early May to August), and recovery stage (August to January). Fully matured oocytes were approximately 250-270 ${\mu}m$ in size. The egg-mass was a hat in shape, and a number of egg capsules were found in an egg-mass. An egg capsule was 0.53-0.57 mm in size. An embryo (veliger larva) hatched from an egg capsule. Percentage of first sexual maturity in females and males were over 50% for individuals that are 40.1-45.0 mm in shell radius, and 100% for those that are over 45.1 mm. The sex ratio of female to male was significantly different from 1:1 $(x^2\;=\;57.22,\;p\;<\;0.05)$.

• Journal of Aquaculture
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• v.6 no.4
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• pp.235-253
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• 1993

The freshwater crab, Eriocheir japonicus inhabits from sub-tropical to temperate zone in Asia. This species belongs to a large size group among freshwater crabs. Common size of this crab is 5-6cm in carapace length and occasionally 7cm in carapace length. This species of crab used to inhabit in estuaries, rivers and inland waters in Korea. However, natural population recently has been rapidly decreased because of pollution and lost their habitats by suburban development. Therefore, development of proper methods of seedling production to increase natural stock became necessity. As parts of achieving this goal, duration from mating to spawning, egg incubation period, and egg development of this species were studied. The influence of temperatures and salinities on the egg incubation and hatching was also investigated. It took 2-8 hours from mating to egg spawning and the spawning lasted 3-9 hours from the first spawning. Egg numbers per female (6cm in carapace length) were 380,000­410,000. Optimum temperature for egg incubation was $17\~23^{\circ}C$ and optimum salinity, $14.0\~31.5\%o$. Incubation period of the eggs at $14^{\circ}C,\;17^{\circ}C,\;20^{\circ}C,\;26^{\circ}C,\;and\;28^{\circ}C$ was 42, 28, 21, 15, and 14 days. respectively. Relation between temperature (X) and incubation days (Y) was LogY = Log 2764.267 - 1.608 LogX. A female can spawn 4-6 times per year by manipulation of environmental conditions. Under the conditions of $18^{\circ}C\;and\;24.5\%o$, it took 6 days up to embryo formation, 18 days up to compound eye formation, 22 days up to abdominal movement, and 25 days up to hatch out as zoea larvae.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.319-326
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• 1996

The objective of this study was to investigate correlation between the morphology by microscopic assessments of surplus blastocysts produced in human IVF program and their cell number obtained by differential labelling method. For these experiments, 76 surplus human blastocysts were obtained from 36 patients on day 5 after IVF, the embryos were classified to early (ErB), early expanding (EEB), middle expanding (MEB), expanded blastocyst (EdB) according to their blastocoel expansion and zona thickness. When the ovum size and zona thickness of the classified blastocysts were measured using micrometer, although the embryos were produced in the same culture condition, there were significant variances in ovum size ($148.8 217.6{\mu}m$) and zona thickness ($1.2-14.4{\mu}m$). Total blastomere cell number counted after hoechst staining was increased by two to three fold during the transition period from ErB ($39.1{\pm}3.6$) to EdB ($(89.6{\pm}3.3)$) stage on day 5 after IVF. ICM ($11.9{\pm}1.8-22.2{\pm}4.3$) and TE ($24.5{\pm}3.6-70.0{\pm}7.7$) cell numbers using differential labelling were also showed the increased pattern according to the developmental level. Especially, EdB which showed poor ICM morphologically also indicated the low ICM cell number after differential labelling. This demonstrated that there is good correlation between the morphological assessment and the cell number. The count of ICM and TE nuclei using differential labelling can be used as an important criterion, if it is accompanied with morphological assessments, in selecting the better embryos for improving the pregnancy rates in human blastocyst transfer program.

• Korean Journal of Animal Reproduction
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• v.22 no.4
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• pp.425-435
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• 1998

To improve the efficiency of nuclear transplantation in bovine, in this study the development in vitro of nuclear transferred (NT) embryos was compared by different activation regimens of the enucleated oocytes. The effect of developmental stage and culture system of donor nuclei on fusion and development in vitro of NT embryos were also evaluated. Oocytes were collected from Hanwoo ovaries obtained from slaughterhouse and matured in Ham's F-10 supplemented with hormones. After 20~22 h maturation, the oocytes were vortexed to be free from cumulus cells and subsequently their nucleus and the first polar body were removed. Enucleated oocytes were divided into 3 groups for activation; the oocytes of group I were activated with ionomycin for 5 min and subsequently incubated in 6-dimetylarninopurine (DMAP) for 4 h, Those of group II were treated with DMAP for 4 h at 39 h after onset of in vitro maturation (IVM) and those of group III were kept in room temperature ($25^{\circ}C$) for 3 h at 39 h after onset of IVM. After in vitro fertilization (IVF) the embryos for muclear donor were cultured either by group culture (20 embryos /50 ${mu}ell$ drop) or individually (1 embryo /50 ${mu}ell$ drop) for 4 day and 5 day. At day 4 and 5 after IVF, blastomeres were separated in calcium-magnesium free medium, and then classified into small (day 5: $\leq$ 38 ${\mu}{\textrm}{m}$, day 4: $\leq$ 46 ${\mu}{\textrm}{m}$) and large (day 5 : $\geq$ 38 ${\mu}{\textrm}{m}$, day 4 ; $\geq$ 46 ${\mu}{\textrm}{m}$). The separated blastomeres were replaced into enucleated and activated recipient cytoplasm. The blastomere-oocyte complexes were fused by electrically. The NT embryos were cultured in TCM-199 containing 10% FCS in 39$^{\circ}C$, 5% $CO_2$ incubator for 7 day. The results obtained were summarized as follows; There were no differences in fusion and development to blastocyst between groups as group I (68%, 10%), group II (75%, 14%) and group III (73%, 9%), respectively. However, the cell number in blastocyst of NT embryos in group III were significantly fewer than in the other groups (P＜0.05). No differences in fusion and development to blastocyst were found between individual or group cultured and between small or large blastomeres of day 4 and day 5 donor embryos. From these results, it was concluded that the combination of ionomycin and DMAP, or treatment of DMAP at 39 h after onset of IVM were useful for the efficient of production of NT bovine embryos, and the individual cultured embryos could be simply used as donor nuclei for NT bovine embryo.

• Journal of Aquaculture
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• v.8 no.3
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• pp.195-207
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• 1995

In order to study the embryonic development and hatching of wild long shanny, Stichaeus grigorjewi, were caught with the gill nets in the East Sea of Korea, and stocked at indoor tanks to induce natural spawning in February 25, 1994 and February 16 to 24, 1995. They were already matured when stocked, and average body length (50.66 cm) and body weight (1,192.74 g) of 57 females and average body length (48.62 cm) and body weight (612.58g) of 43 males were recorded. Before stocking, they were inserted with identification tags(ID tags) in the dorsal muscle, and spawning was traced by the portable reader (Destron/lDl Ltd.) Forty females among 57 spawned successfully in the average of 4 days after stocking. Females spawned almost all eggs contained in the ovaries at one time in the form of an egg mass and averaging 227,200 eggs Per egg mass. The egg mass was oval in shape, translucent milky in color, 20.32cm long axis and 14.57cm short axis in size, and 803.7g in weight. Male parents guarded their egg masses and circulated water with the tail part of the body. Fertilized egg was spherical in shape, and their average diameter was 1.54 mm. Each egg had a containing single oil globule, and it's average diameter was 0.37 mm. The average water temperature was $13.2^{\circ}C$ and incubation times after fertilization were 5 hours 25 minutes up to 2-cell stage, 13 hours up to morula stage, and 66 hours 35 minutes up to embryo formation stage. Hatching rate was approximately 10 percent in 368 hours 50 minutes after fertilization, and approxionateoly 90 percent of eggs were hatched in 425 hours 30 minutes after fertilization.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.52 no.4
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• pp.429-438
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• 2007

Early growth vigor(EGV) is one of the physiological characteristics that may contribute to the increase of genetic yield potential and radiation use efficiency by closing the canopy earlier. To estimate the genotypic variation of EGV, determine the relationships among the related traits, and identify the rapidly growing genotypes and indirect indicator for selection in breeding program, the evaluation of EGV and EGV-related traits was conducted for a total of 140 rice varieties consisting of 101 Korean, 25 Northern China and 14 IRRI-bred rice varieties in a serial sowing experiment in plastic rain shelter and plastic-covered nursery bed in 2003. EGV defined as the amount of leaf area and/or dry weight produced early in the season and the EGV-related traits such as length and breadth of the $2^{nd}\;and\;3^{rd}$ leaves showed highly significant positive correlation with the embryo and seed weight. Especially, the genotypic variation in the length of the third leaf was explained over 90% of genotypic variation in the seed weight. Owing to a large effect of seed size on EGV and its related traits, vigor measurements were adjusted based on their linear or exponential relationships with seed weight for excluding the seed weight effect. EGV and its related-traits adjusted for seed weight also showed big variation among genotypes. Increased EGV was genetically correlated with increases in breadth and length of early leaves. The broad-sense heritability for EGV was significantly high(81%), but lower than those of leaf breadth(90% for the $2^{nd}$ leaf and 93% for the $3^{rd}$ leaf) and length(87% for the $2^{nd}$ leaf and 89% for the $3^{rd}$ leaf). Significantly positive genetic correlations were found between EGV and the breadth and length of early leaves. The high heritability of early leaf breadth and length coupled with their strong genetic correlation with EGV indicated that the breadth and length of the $2^{nd}\;and\;3^{rd}$ leaf would be used as good indirect indicators for EGV selection in rice breeding program.